Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
  • C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
  • C12N11/093—Polyurethanes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P7/00—Preparation of oxygen-containing organic compounds
  • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
  • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P7/00—Preparation of oxygen-containing organic compounds
  • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
  • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
  • C12P7/06—Ethanol, i.e. non-beverage
  • C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P7/00—Preparation of oxygen-containing organic compounds
  • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
  • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
  • C12P7/16—Butanols
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P7/00—Preparation of oxygen-containing organic compounds
  • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
  • C12P7/26—Ketones
  • C12P7/28—Acetone-containing products
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
  • Y02E50/00—Technologies for the production of fuel of non-fossil origin
  • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

• the present description relates to a process for the production of alcohols by fermentation of a sweet fluid.
• Alcohols from fermentation processes are among the most promising substitutes for petrochemical derivatives.
• ABE Acetone - Butanol - Ethanol
• IBE fermentation carried out by microorganisms belonging to the genus Clostridium, is one of the oldest fermentations to have been industrialized (early 20th century) and has since been widely studied. More recently, the IBE fermentation (Isopropanol - Butanol -
• Ethanol producing a mixture of isopropanol, butanol and ethanol and also produced by microorganisms belonging to the genus Clostridium, has been the subject of numerous studies.
• a continuous process with cells suspended in a homogeneous reactor can also be envisaged. But productivity is also quite low and can hardly be increased significantly.
• One of the major technical problems is the concentration of cells in the fermentation medium, mainly controlled by the dilution rate applied in the process. This cannot be raised to avoid cell washing ("wash out” according to English terminology) in the fermenter. For these reasons, for a few years, a great interest is carried by the methods aiming at a strong retention of the microbial biomass. There are two ways: “cell immobilization” and cell "recycling” with retention using filter membranes.
• EPS polysaccharides excreted by their own microorganisms
• a first object of the present description is to provide a fermentation process of the IBEA type (Isopropanol - Butanol - Ethanol - Acetone) in a reactor in which the rate of hydraulic dilution is different from the dilution rate of the active biomass.
• IBEA type Isopropanol - Butanol - Ethanol - Acetone
• a method capable of at least partially fixing the bacterial biomass by adsorption in the form of biofilm in the fermentation reactor, improving the volume productivity is described below.
• the abovementioned object as well as other advantages, are obtained by a process for the production of alcohols, in which a sweet fluid is introduced into a fermentation reactor to produce a fermentation must enriched in isopropanol, butanol, ethanol and acetone with respect to the sweet fluid, the fermentation reactor comprising a biomass produced by a strain belonging to the genus Clostridium supported (ie immobilized) on a solid support comprising a polyurethane foam.
• the fermentation must comprises an addition of at least 0.2 g / L of isopropanol, of at least 0.2 g / L of butanol, of at least 0.2 g / L of ethanol and at least 0.2 g / L of acetone, relative to the sweet fluid.
• the fermentation must comprises a contribution of at least 0.2 g / L of isopropanol, of at least 0.2 g / L of butanol, of less than 0.2 g / L of ethanol and at least 0.2 g / L of acetone, relative to the sweet fluid.
• the fermentation must 0 comprises a contribution of at least 0.2 g / L of isopropanol, of at least 0.2 g / L of butanol, of less than 0.2 g / L of ethanol and of less 0.2 g / L of acetone, compared to the sweet fluid.
• the fermentation must comprises an addition of minus 1 g / L of isopropanol, at least 2 g / L of butanol, relative to the sweet fluid.
• the fermenting must comprises a contribution of at least 2 g / L of isopropanol, of at least 4 g / L of butanol, relative to the sweet fluid.
• the fermentation must comprises an addition of at least 3 g / L 5 of isopropanol, of at least 6 g / L of butanol, relative to the sweet fluid.
• the fermentation must comprises a contribution of at least 4 g / L of isopropanol, of at least 8 g / L of butanol, relative to the sweet fluid.
• the fermentation must comprises an addition of at least 10 g / L of isopropanol, of at least 20 g / L of butanol, relative to the sweet fluid.
• the fermentation must comprises an addition of at least 15 g / L of isopropanol, of at least 30 g / L of butanol, relative to the sweet fluid.
• the fermentation must comprises an intake of at least 0.4 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1 (eg 0.01 to 0 , 99).
• the fermenting must comprises a contribution of at least 3 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio can vary from 0 to 1 (eg 0.01 to 0.99).
• the fermentation must comprises an intake of at least 6 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1 (eg 0.01 to 0 , 99).
• the fermentation must comprises an intake of at least 90 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1 (eg 0.01 to 0 , 99).
• the fermentation must comprises an intake of at least 12 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1 (eg 0.01 to 0 , 99).
• the fermentation must comprises an intake of at least 30 g / L 5 of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1
• the fermentation must comprises an intake of at least 60 g / L of isopropanol + butanol, the isopropanol / (isopropanol + butanol) ratio being able to vary from 0 to 1 (eg 0.01 to 0 , 99).
• the production process according to the first aspect also makes it possible to maintain better stability of the microorganisms (eg maintenance of performance over time).
• the fermentation products, and particularly the alcohols (eg butanol) have an inhibiting effect on the microorganism
• the production process according to the first aspect reduces at least in part the negative effects of these inhibitor products on the biological activity present in the environment.
• the sweet fluid is introduced continuously into the fermentation reactor.
• the polyurethane foam comprises at least one of the following characteristics:
• - volume cavities ie., pores or cells
• equivalent sphere diameter is between 0.1 and 5 mm, preferably between 0.25 mm and 1.1 mm, preferably between 0.55 and 0, 99 mm, and
• an apparent density ie, mass on apparent volume measured in the air between 10 and 90 g / L, preferably between 10 and 80 g / L, preferably between 15 and 45 g / L, such as between 20 and 45 g / L.
• the fermentation is carried out at a dilution rate (defined as the charge flow ratio (liquid volume of the sweetened fluid) to be converted relative to the liquid volume of the fermentation reactor) of between 0.04 h 1 and 1 h 1 , preferably between 0.08 h 1 and 0.5 h 1 , such as between 0.12 h 1 and 0.3 h 1 .
• a dilution rate defined as the charge flow ratio (liquid volume of the sweetened fluid) to be converted relative to the liquid volume of the fermentation reactor
• the fermentation reactor comprises between 10% and 90%, preferably between 20% and 50%, preferably between 20% and 40% (eg 25-30%) in apparent volume of solid support by relative to the total volume of the fermentation reactor.
• the solid support is at least partially, preferably completely immersed, in the reaction medium.
• the solid support is traversed by natural or forced convection with a flow of sweet fluid (eg descending, rising, or radial fluid circulation, optionally in forced convection (eg radial turbine of the Rushton or axial type or through a support)).
• the fermentation reactor is an upward or downward or radial fluid circulation reactor and optionally with countercurrent gas evolution.
• the biomass is produced by (and / or comprises) a microorganism belonging to the genus Clostridium and capable of producing mixtures of the IBEA type (eg Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium saccharobutylicum. Clostridium tyrobutyricum, C. saccharoperbutylacetonicum, C. butylicum 5 and other Clostridium sp).
• the microorganism employed may or may not be genetically modified.
• Clostridium species known as “solvents” can be used.
• the strains preferably used are those belonging to the species C. beijerinckii or C. acetobutylicum. They can be genetically modified strains or not.
• a genetically modified strain corresponds to a strain whose genetic material (DNA) has been modified compared to an initial strain. Genetic modifications are carried out using genetic tools well known to those skilled in the art (cf. Pyne et al Biotech Adv 2014 32 (3): 623-41 and Wasels et al J. Microbiol Methods 2017 140: 5-1 1).
• the genetic modifications may correspond to modifications of the own genomic content of the strain used in order to improve its performance for the production of Isopropanol / Butanol / Ethanol or its capacity to modify the selectivity towards isopropanol or n-butanol. Genetic modifications may also correspond to the integration of one (or more) genetic material (s) making it possible to improve the performance or the selectivity towards isopropanol or n-butanol of the Clostridium strains used in the process.
• the biomass produced by the strain belonging to the genus Clostridium comprises a bacterium 5 genetically modified or not and belonging to the species Clostridium beijerinckii and / or
• the sweet fluid comprises an aqueous solution of sugars derived from C5 and / or C6 lignocellulose, and / or sugars derived from saccharifous plants (eg glucose, fructose and sucrose), and / or sugars from starchy plants (eg dextrins, maltose and other oligomers, even starch).
• the aqueous solution comprises from 20 to 800 g / L (eg from 20 to 500 g / L) of sugar.
• the sweet fluid is produced from a load of biomass.
• the biomass load comes from the treatment of a renewable source.
• the renewable source comprises lignocellulosic biomass (eg woody substrates, such as softwoods and hardwoods (for example softwoods such as spruces or pines, or hardwoods such as eucalyptus), agricultural products (eg straw) or those of industries generating lignocellulosic waste (food industries, paper mills)) and / or plants from dedicated cultures (eg miscanthus, Panicum virgatum (panic erect or switchgrass)), and / or sugar plants, such as sugar beet, sugar cane, Jerusalem artichoke and / or starchy plants (eg corn and wheat) and / or tubers (eg cassava, Jerusalem artichoke and potato).
• the renewable source further comprises a lignocellulosic biomass of products and residues from the paper industry and transformation products of lignocellulosic materials. 5
• the biomass load comprises approximately 35 to
• the solid support is placed inside the fermentation reactor before and / or after the immobilization (e.g. by adsorption) of the biomass produced by the strain belonging to the genus Clostridium on the solid support.
• the biomass produced by the strain belonging to the genus Clostridium is immobilized on the solid support in a secondary reservoir, and the solid support supporting the biomass produced by the strain belonging to the genus Clostridium (also named below) bacterial biomass) is introduced into the fermentation reactor. It is possible to immobilize the bacterial biomass inside a secondary tank operating in a fast loop ("in stream" according to English terminology) relative to the fermentation reactor. It is in particular possible to implement a batch stage in the process of the present description, for example for a period of 4-5 hours, for example before the continuous introduction of the sweet fluid into the fermentation reactor.
• the reaction medium which contains / submerges the solid support undergoes an inoculation rate with an inoculation solution of bacterial biomass (eg with cells at substantially maximum growth rate) of between 0.5 % and 20% by volume, preferably between 1% and 20% by volume relative to the total volume of the reaction volume (eg 10% by volume).
• the solid support is at least partially, preferably completely immersed, in the inoculation solution during said “batch” step.
• the bacterial biomass is immobilized on the solid support in the form of a biofilm (ie microbial group which can either be attached to a solid surface (organic or inorganic), or form flocs or aggregates in isolation , especially by auto granulation, on the solid surface).
• a biofilm ie microbial group which can either be attached to a solid surface (organic or inorganic), or form flocs or aggregates in isolation , especially by auto granulation, on the solid surface).
• the solid support is introduced into the fermentation reactor in the form of one or more blocks.
• the solid support e.g. in the form of a single block
• the block has a diameter substantially equal to the internal diameter of the fermentation reactor.
• the block and the fermentation reactor are described herein as straight circular cylinders, it is understood that the block and the fermentation reactor can be of any shape.
• the block is centered or eccentric relative to the main (vertical) axis of the fermentation reactor, or attached to a radial wall of the fermentation reactor.
• the solid support is introduced into the fermentation reactor in the form of a net or a container with a mesh comprising a plurality of cubes, parallelepipeds or any other 3-dimensional shapes ("chips" according to Anglo-Saxon terminology) of polyurethane foam, for example at least one dimension of which is at least 3 mm.
• the net or the container with a mesh is a straight circular cylinder having a diameter less than or substantially equal to the internal diameter of the fermentation reactor.
• the net or the container with a mesh has a diameter substantially equal to the internal diameter of the fermentation reactor.
• the solid support forms a fluidized bed or a fixed bed in the fermentation reactor.
• the solid support forms a fluidized bed maintained in immersion, at least partial and preferably total, by a grid.
• the solid support is agitated (eg mechanically).
• the solid support is immobilized inside a net or a container with a concentric mesh, for example concentric with the stirring axis (eg use of a Robinson type reactor Mahoney).
• the solid support e.g. block (s), net, container with mesh
• the sweet fluid is introduced directly above or below the block (s), the net or the mesh.
• the fermentation is an anaerobic (0 strict) fermentation, such as under a supply of inert gas (e.g. under nitrogen).
• inert gas e.g. under nitrogen
• the fermentation is carried out at a temperature between 28 ° C and 40 ° C, preferably between 30 ° C and 37 ° C (eg 36 ° C), and / or at a pressure between about 0.1 MPa and 0.15 MPa (ie, atmospheric pressure + water heights). 5 According to one or more embodiments, the fermentation is carried out continuously for a duration of at least 250 hours, preferably at least 500 hours without any superior limitation (e.g. 5000h).
• a fermentation reactor (2) comprising a biomass produced by a strain belonging to the genus Clostridium supported on a solid support (9) comprising a polyurethane foam .
• At least part of the fermentation must obtained at the outlet of the fermentation reactor is recycled at the inlet of the fermentation reactor. It is in particular possible to reach linear speeds independent of the overall residence time 5.
• Figure 1 is a schematic view of a process for the production of alcohols according to embodiments of the present description.
• FIG. 2 is a schematic view of a solid support whose diameter is substantially equal to the internal diameter of a fermentation reactor according to embodiments of the present description.
• Figure 3 is a schematic view of solid supports centered, eccentric or attached to the walls of fermentation reactors according to embodiments of the present description.
• FIG. 4 is a schematic view of solid supports according to embodiments of the present description, comprising elements of polyurethane foam confined in a net or a container with a mesh.
• FIG. 5 is a schematic view of a solid support forming a fluidized bed contained in immersion in a fermentation reactor according to embodiments of the present description.
• FIG. 6 shows a schematic view of a process for the production of alcohols according to embodiments of the present description, further comprising a step of finishing fermentation.
• FIG. 7 shows the evolution of the volume productivity in IBEA of a reference process as a function of the imposed dilution rate.
• the sweet fluid is the sweet fluid
• the sweet fluid comprises an aqueous solution of sugars derived from C5 and / or C6 lignocellulose, and / or sugars derived from saccharifous plants (eg glucose, fructose and sucrose), and / or sugars from starchy plants (eg dextrins, maltose and other oligomers, even starch).
• the aqueous solution of sugars C5 and / or C6 comes from the treatment of a renewable source.
• the renewable source is of the lignocellulosic biomass type which can in particular include woody substrates (eg hardwoods and conifers), agricultural by-products (eg straw) or those of industries generating lignocellulosic waste (eg food industry, paper mills).
• the renewable source can also come from sugar plants, such as sugar beet and sugar cane or starchy plants such as corn and wheat.
• the aqueous solution of C5 and / or C6 sugars can also come from a mixture of different renewable sources.
• the bacterial biomass is mainly adsorbed in the form of biofilm on a solid support.
• the bacteria are strains belonging to the species Clostridium beijerinckii and / or Clostridium acetobutylicum.
• the bacteria used in the process can be genetically modified strains or not and naturally producing isopropanol and / or Clostridium strains naturally producing acetone genetically modified to make them produce isopropanol.
• the solid support includes a polyurethane foam.
• Polyurethane foam is particularly advantageous because it not only gives access to the production of IBEA type mixtures, but it also gives access to continuous type production by immobilizing the bacterial biomass. Indeed, the Applicant has demonstrated that polyurethane foam is capable of fixing bacteria of the genus Clostridium from sufficiently large (ie., beyond the dilution rate causing cell washing) allowing continuous production of IBEA type mixtures.
• the polyurethane foam is adapted to be immobilized by immersion in a reactor.
• the polyurethane foam has:
• 5 - volume cavities ie, pores or cells whose equivalent sphere diameter is between 0.1 and 5 mm, preferably between 0.25 mm and 1.1 mm, preferably between 0.55 and 0, 99 mm, and / or
• an apparent density ie, mass on apparent volume measured in the air between 10 and 90 g / L, preferably between 10 and 80 g / L, preferably between 0 15 and 45 g / L, such as between 20 and 45 g / L or between 25 and 45 g / L.
• the equivalent sphere diameter of the volume cavities can in particular be obtained by analysis with a X-ray microscope (eg HR tube 70kV 200 microA Medium focal point; Varian pixel detector: 6 microns; acquisition time: 2 hours) of a sample (eg 7 mm x 7mm x 15 5 mm) and reconstruction of a representative volume of the foam (eg reconstructed volume 5 mm x 5 mm x 5 mm with a voxel size of 6 microns) with the assumption of spherical cells.
• a X-ray microscope eg HR tube 70kV 200 microA Medium focal point; Varian pixel detector: 6 microns; acquisition time: 2 hours
• a sample eg 7 mm x 7mm x 15 5 mm
• reconstruction of a representative volume of the foam eg reconstructed volume 5 mm x 5 mm x 5 mm with a voxel size of 6 microns
• Diameter measurements were made by 3D image analysis with Avizo software from 3D volumes acquired by X-ray microscope.
• the cells were closed artificially0 by image analysis so as to estimate the volume and then the diameter.
• the diameter of a given cell is assimilated to that of a sphere of the same volume.
• the different stages of image analysis are as follows:
• FIG. 1 represents a diagram of production of a mixture of alcohols from a substrate of the lignocellulosic biomass type.
• the sweet fluid comprising for example C5 and / or C6 sugars is introduced via line 1 into a fermentation reactor 2 to undergo a fermentation step.
• the sweet fluid is brought into contact with the bacterial biomass supported on a solid support comprising a polyurethane foam.
• Fermentable sugars eg C5 and / or C6 sugars
• alcohols and / or 0 solvents by microorganisms to produce a (first) fermentative must (or juice or wine), in particular enriched in isopropanol, butanol, ethanol and acetone versus sweet fluid.
• the fermentation step in the fermentation reactor 2 can be carried out at a temperature between 28 ° C and 40 ° C, preferably between 30 ° C and 37 ° C, so that the fermentative must include products of the IBEA type fermentation reaction, for example isopropanol which is then discharged via line 3.
• the fermentation must is introduced via line 3 into a separation unit 4 (optional) making it possible to separate and extract the compounds of interest from the fermentation must, the latter being evacuated via line 5.
• the residues of the separation commonly called vinasses, are evacuated from the separation unit 4 through the conduit 6. Les0 vinasses are generally composed of water as well as any liquid or solid product that is not converted or not extracted during the preceding steps.
• the separation unit 4 can carry out one or more distillations, and optionally a separation of the solid and / or suspended matter, for example by centrifugation, decantation and / or filtration.
• bacterial biomass can be immobilized on a solid support directly in the fermentation reactor 2 or indirectly in a secondary tank 7 (optional), operating for example in "in stream” mode0 with respect to the fermentation reactor 2.
• the solid support thus in charge of Bacterial biomass can then be introduced into the fermentation reactor, for example via line 8 or any other means.
• the solid support forms a fluidized bed or a fixed bed.
• the liquid surface speed of the sweet fluid is greater than the minimum fluidization speed.
• the liquid surface speed of the sweet fluid is modified (e.g. increased) as a function of the evolution of the density difference which takes place during fermentation. For example, as the biofilm is formed on the solid support, the density of the solid support may vary (eg increase), giving rise to evolving hydraulic regimes (eg different optional recycling rates at the start of fermentation and at the end of fermentation).
• a solid support comprising a loose or structured stack of 5 particles of polyurethane foam can be envisaged, with or without mechanical agitation
• the fermentation medium crosses the solid bed in an upward or downward current (“upflow” or “downflow” according to English terminology).
• upflow or “downflow” according to English terminology
• a system0 allowing a gaseous release against the current can be provided.
• a radial circulation can also be envisaged in the fermentation reactor 2, for example in the case where mechanical agitation is applied to the center of the fermentation reactor 2 (e.g. radial turbine of the Rushton type).
• the solid is immobilized inside a basket concentric with the stirring axis 5, making it possible in particular to control the speeds of the reaction medium and the hydrodynamics imposed around the latter (eg Robinson Mahoney type reactor).
• the solid support is partially or completely submerged, in particular to increase the formation of biofilms and improve performance.
• the solid support is introduced in the form of a single block, for example in the form of a cylinder whose diameter is less than or substantially equal to the internal diameter of the fermentation reactor 2.
• the diameter of the solid support 9 is substantially equal to the internal diameter of the fermentation reactor 2.
• the block can thus correspond to a filtering medium within which the biofilms will develop.
• the solid support 9 as shown in FIG. 02 can cause a slight overpressure in the free liquid phase, at the lower level. It should be noted that if the gaseous evolution is significant during fermentation, a gas pocket can form under the solid support and preferential passages within the solid support 9 can be generated during the evacuation of the gas.
• the solid support block 5 can be centered, eccentric or attached to a wall of the fermentation reactor 2.
• the solid support 9 does not in any way disturb the circulation of the liquid entering or leaving the process, in particular when operated continuously.
• the possible presence of insolubles such as those from large grains does not pose any problems.
• the flow of sweet fluid arriving via the conduit 1 can also be introduced to the right of the solid support blocks 9, for example when these are flush with the surface of the reaction medium of the fermentation reactor 2.
• the solid support is flush with the surface of the reaction medium at the entry of the sweet fluid, the medium is locally less concentrated in alcohol and the growth of bacteria is favored.
• the solid support comprises a net or a container with a mesh 10 comprising cubes or parallelepipeds or other elements of any 3-dimensional shape (eg polyhedra) of large or small size (eg at least one dimension between 3 mm and 10 m, such as from 2 cm to 1 m), as shown in Figure 4, the elements being made of polyurethane foam.
• the net or the container with mesh 10 forms a cylinder whose diameter is less than or substantially equal to the internal diameter of the fermentation reactor 2.
• the diameter of the net or of the container with mesh 10 is substantially equal to the internal diameter of the fermentation reactor 2. It may be that gas emissions tend to raise the solid support 9.
• a perforated plate, a simple net or a grid 1 1 may be sufficient to keep the solid support, for example in motion, in the fermentation reactor 2.
• the fermentation reactor 2 has an upward fluid circulation.
• the direction of circulation of the sweet fluid can be descending. It is also understood that the direction of circulation can be generally rising or falling (seen from the outside of the fermentation reactor) and radial inside the fermentation reactor 2.
• the process for the production of alcohols can use a finishing reactor 12 called “finisher” (optional).
• the fermentation must withdrawn from the fermentation reactor 2 through the conduit 3 is introduced into the finishing reactor 12 adapted to produce a second fermentation must enriched in IBEA compared to the fermentation must withdrawn from the fermentation reactor 2.
• the second fermentation must is then withdrawn from the finishing reactor 12 and evacuated via line 6 and introduced into the separation unit 4 (optional) making it possible to separate and extract the compounds of interest from the fermentation must, the latter being evacuated via line 5.
• the finishing reactor 12 is preferably without PU foam.
• the finisher is preferably homogeneous and aims to exploit the IBEA title potential of the Clostridium strain as well as possible, by allowing an extended residence time, and to add an amount of carbonaceous substrate which corresponds to the needs of the strain.
• the finishing reactor 12 makes it possible in particular to guarantee an exhaustion of the dares.
• the process for producing alcohols also puts in place a step of partial recovery of the IBEA compounds produced present in the first and / or the second fermenting must.
• This IBEA compound extraction step uses a stripping step with a pressurized gas sent to the fermentation reactor 2 and / or the finishing reactor 12 in order to entrain the alcohols present in the aqueous phase.
• the stripping gas is a gas produced directly by fermentation and which has been previously stored (by methods known to those skilled in the art) before its implementation.
• the stripping gas typically comprises carbon dioxide and optionally hydrogen.
• This gas stripping step advantageously makes it possible to control the alcohol content during fermentation. present in the environment in order to limit the phenomena of inhibition of microorganisms which appear when the alcohol content reaches a critical value.
• the gas stripping step can either be carried out continuously or discontinuously.
• the fermentation gas flow rate relative to the fermenter volume is for example between 0.5 and 2.5 l / l / min, preferably between 0.7 and 1.1 l / l / min.
• the recovery process can also be implemented so that the gas stripping step is carried out in a finisher 12 containing an organic solvent immiscible with water, the solvent forming a organic phase supernatant above the fermentation must.
• the solvent will also be chosen so as to be biocompatible with the microorganism.
• the stripping gas is thus injected into the fermentation must so as to entrain the alcohols produced in the supernatant organic phase and in such a way that part of the alcohols are transferred to the organic phase when the stripping gas passes through said organic phase.
• a first batch stage (duration 4-8 hours) corresponding to the lag time (“lag time” according to English terminology) and start of exponential growth (accompanied by generation of fermentation gas);
• FIG. 7 shows the evolution of the volume productivity r in IBEA g / Lh as a function of the dilution rate D imposed in the bioreactor (h 1 ).
• the dilution rate D is defined as the volume flow entering the reactor divided by the volume of the last.
• this parameter can be considered both as the inverse of the residence time for the fluid and for the microorganisms. Consequently, a cell washing phenomenon appears beyond a certain dilution rate, leading to a loss in volume productivity.
• the critical dilution rate is between 0.04 and 0.06 h 1 , when the maximum productivity reaches a value of approximately 0.45 g / Lh of IBEA.
• Example 1 according to the present description: Batch mode test with polyurethane foam
• bioreactors are filled with 20 mL of fermentation medium, which has previously been placed under anaerobic conditions in order to guarantee the absence of oxygen. 40% of solid volume (apparent volume) relative to the total volume of the bioreactor is introduced into each bioreactor.
• the initial glucose is fixed at 90 g / L, the seeding rate is 10% (liquid vol) (same inoculum for all fermentations) with cells at the maximum growth rate.
• the microorganism used is Clostridium beijerinckii DSM 6423. All the bioreactors are introduced into an anaerobic jar, and brought to temperature (36 ° C), for a fixed duration of 12 days. The pressure is substantially atmospheric + the water height of the bioreactor. Then, the final fermentation must is analyzed, as well as the solids supporting biofilm. Each operating condition is carried out in triplicate to ensure the repeatability of the experiments.
• the titer is quantified from the fermentation yield which is considered invariable and the glucose consumption on each test.
• batch fermentations with Mousse 1 and Mousse 2 produced 17.5 g / L and 15.2 g / L of IBEA respectively, a titer in both cases higher than that obtained with control fermentation (13, 3 g / L IBEA)
• Example 2 continuous test with polyurethane foam
• the process is implemented experimentally according to embodiments of the present description, by means of two fermentations in continuous mode with immobilization on solid supports of the foam type.
• polyurethane Foam 1, Foam 2 with different physical and structural characteristics.
• the main characteristics of these two foams are explained in Table 1.
• the filling is carried out in bulk mode, with cubes of dimensions 3 mm x 5 mm x 5 mm.
• a new period of time corresponding to at least three times the residence time is expected with each new set point for stabilization.
• the final fermentation wort is taken sterile from the column and analyzed for glucose and major metabolites (i.e. isopropanol, butanol, ethanol and acetone).
• Table 3 shows the evolution of the fermentation, as a function of the imposed dilution rate, of the percentage of glucose consumption (% ), de0 the total IBEA content (g / L), and the volume productivity (g / Lh in IBEA). Compared to the conditions of suspended cells (reference example), an increase in productivity is observed in the case where the cells are immobilized on Moss 1 (factor 6) or Mousse 2 (factor 3).
• the bacterial biomass used in the process according to the present description may correspond to another strain than Clostridium beijerinckii DSM 6423.
• the polyurethane foams according to the present description may be other than those described in Table 1.
• IBEA contents and improved volume productivity can be obtained by means of inoculation rate, sugar rate, dilution rate, temperatures, pressures, agitation, durations, etc. other than those shown in the examples. 0
• all the embodiments described above can be combined with one another.
• all of the features of the embodiments described above can be combined with or replaced by other features of other embodiments. 5
• the process is implemented experimentally according to embodiments of the present description, by means of two fermentations in continuous mode with immobilization on solid supports of the polyurethane foam type.
• the filling is carried out in bulk mode, with cubes of dimensions 10 mm x 10 mm x 7 mm.
• the foams have a macro-pore size of about 1 mm.
• the two fermenters are in the form of a glass column of 250 ml of useful volume.
• the conditions for filling the fermenters are explained in Table 4.
• a recirculation loop is arranged between the inlet and the outlet of the reactor to maintain good homogenization within the fermenter.
• the liquid leaves by overflow.
• the entire fermentation system was previously put under anaerobic conditions by nitrogen purge to guarantee the absence of oxygen.
• the glucose concentration in the feeder is fixed at 60 g / L.
• the fermenter is inoculated at 10% by volume with cells at the maximum growth rate relative to the total volume of the fermentation medium.
• the microorganism used is Clostridium beijerinckii DSM 6423.
• the system is temperature controlled at 34 ° C without agitation other than recirculation.
• the pressure is substantially atmospheric. 0
• the test takes place in 2 stages:
• the dilution rate is continuously increased as a function of the solvent concentrations.
• the fermentation must is removed sterile and analyzed for glucose and main metabolites (i.e. isopropanol, butanol, ethanol and acetone). .
• the pH is also measured. 0
• the two fermenters are operated over a period of 912 h.
• the dilution rate varies between 0.02 h-1 and 0.23 h-1.
• the total IBEA content varies between 8 and 16 g / L.
• the maximum volume productivity for the two fermenters is 1.5 g / L. h in IBEA.
• the best performance over the entire test gives a maximum volume productivity of 2.44 g / L / h in IBEA for fermenter 1 and 2.24 g / L / h 5 in IBEA for fermenter 2.
• Example 4 continuous test with polyurethane foam in a stirred reactor
• a continuous test is carried out experimentally with cells immobilized on a support of the polyurethane foam type.
• a bioreactor with 5 L0 of total volume is filled with 2 L of fermentation medium.
• the initial glucose is fixed at 60 g / L, and the inoculum is 0.2L, i.e. an inoculation rate at 10% by volume relative to the volume total fermentation medium with cells at maximum growth rate, after having undergone a 1 hour nitrogen purge in order to ensure anaerobiosis (strict) from the start of the purge test nitrogen is maintained during the preliminary batch stage (7 hours).
• the microorganism used is Clostridium beijerinckii DSM 6423.
• the temperature is fixed at 5 34 ° C. Mechanical agitation varies between and 60 and 170 rpm.
• the pressure is substantially atmospheric + the water height of the bioreactor.
• the test takes place in 2 stages:
• the dilution rate is continuously increased as a function of the solvent concentrations.
• the fermentation wort is removed sterile 5 and analyzed for glucose and main metabolites (i.e. isopropanol, butanol, ethanol and acetone). The pH is also measured.
• the two fermenters are operated for over a period of approximately 765 h.
• the dilution rate varies between 0.02 h-1 and 0.2 h-1.
• the total IBEA content varies between 5 and 16 g / L.
• the maximum volume productivity obtained for this fermentation is 1.6 g / L. h en0 IBEA.

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